PROJECT SUMMARY: FLOW CYTOMETRY AND CELLULAR IMAGING FACILITY (FCCIF) The Flow Cytometry and Cellular Imaging Facility (FCCIF) was established to provide access to state-of-the-art equipment for single-cell analysis and isolation, and to provide expertise in cell sorting, analytical flow cytometry, cellular imaging, and custom monoclonal antibody-fluorochrome conjugations. Core personnel are available to design and execute experiments and analyze the results using a variety of commercially available or Core- designed custom software packages. The Core is comprised of 2 independently managed sites: one on the North Campus (NC) and the other on the South Campus (SC). Dr. Michael Andreeff is the director of the FCCIF; Dr. Jared K. Burks is co-director of the NC site, and Dr. Karen Clise-Dwyer is co-director of the SC laboratory. The FCCIF-NC specializes in fluorescence-activated cell sorting, sorting, mass cytometry (CyTOF -cytometry by time of flight), and cellular imaging. A major investment has been made in mass cytometry; the Core is now part of The University of Texas System Proteomics Network and has 2 CyTOFs and a Hyperion mass cytometer funded by both MD Anderson Cancer Center and The University of Texas System. The FCCIF-SC specializes in highly multi-parametric fluorescence-based analytical flow cytometry and cell sorting, and offers imaging flow cytometry using an Amnis ImageStream system. Over the past 6 years, the institution has provided $695,000 in renovations and $4,512,156 in funds for capital equipment. The FCCIF now uses 27 major instrument systems (NC: 14; SC: 13) and over the past 6 years has provided service to 404 cancer center members in support of all 16 CCSG programs as well as numerous MD Anderson Moon Shot programs and platforms, P01s, R01s, and P50 SPOREs. Cancer center members with peer-reviewed funding account for 92% of use, and 28% ($467,180) of the total Core budget is requested from the CCSG. In the current grant cycle, the FCCIF has provided 95,375 hours of service, and grant Yr42 had a 35% increase in use over that in Yr37 and a 63% increase over the average in the previous grant period. Since the previous grant period, the FCCIF has supported 589 publications, with 421 (71%) appearing in journals with IF >5 and 167 (28%) in journals with IF >10, including Science, Nature, Nat Med, Nat Immunol, Cell, Immunity, Cancer Cell, and Cell Stem Cell. The FCCIF specific aims are: Aim 1) Collaboration: To provide the MD Anderson research community with unparalleled expertise to address important research hypotheses using robust, state-of-the-art flow cytometry and cellular imaging techniques; Aim 2) Innovation: To develop and validate new methods in the fields of cytometry, cell sorting, and cellular imaging; and Aim 3) Education: To educate users in applicable methodology and best practices in data collection and analysis to facilitate research rigor and reproducibility.